Determination of the intramolecular disulfide bond arrangement and biochemical identification of the glycosylation sites of the nonstructural protein NS1 of Murray Valley encephalitis virus

The 12 cysteine residues in the flavivirus NS1 protein are strictly conserved, suggesting that they form disulfide bonds that are critical for folding the protein into a functional structure. In this study, we examined the intramolecular disulfide bond arrangement of NS1 of Murray Valley encephalitis virus and elucidated three of the six cysteine-pairing arrangements. Disulfide linkages were identified by separating tryptic-digested NS1 by reverse-phase high pressure liquid chromatography and analysing the resulting peptide peaks by protein sequencing, amino acid analysis and/or electrospray mass spectrometry. The pairing arrangements between the six amino-terminal cysteines were identified as follows: Cys(4)-Cys(15), Cys(55)-Cys(143) and Cys(179)-Cys(223). Although the pairing arrangements between the six carboxyterminal cysteines were not determined, we were able to eliminate several cysteine-pairing combinations. Furthermore, we demonstrated that all three putative N-linked glycosylation sites of NS1 are utilized and that the Asn(207) glycosylation site contains a mannose-rich glycan.

Antibody-dependent enhancement of Murray Valley encephalitis virus virulence in mice

Enhancement of flavivirus infection in vitro in the presence of subneutralizing concentrations of homologous or heterologous antiserum has been well described. However, the importance of this phenomenon in the enhancement of flavivirus infection in vivo has not been established. In order to study antibody- mediated enhancement of flavivirus infection in vivo, we investigated the effect of passive immunization of mice with Japanese encephalitis virus (JE) antiserum on the outcome of infection with Murray Valley encephalitis virus (MVE). We show that prior treatment of mice with subneutralizing concentrations of heterologous JE antiserum resulted in an increase in viraemia titres and in mortality following challenge with wild-type MVE. Our findings support the hypothesis that subneutralizing concentrations of antibody may enhance flavivirus infection and virulence in vivo. These findings are of potential importance for the design of JE vaccination programs in geographic areas in which MVE co-circulates. Should subneutralizing concentrations of antibody remain in the population following JE vaccination, it is possible that enhanced disease may be observed during MVE epidemics.

Northern wing, General Purpose North 3, University of Queensland, St Lucia

As seen from Blair Road.

Northern wing, General Purpose North 3, University of Queensland, St Lucia

As seen from Balir Road.

West elevation, General Purpose North 3, University of Queensland, St Lucia

Formal elevation facing University of Queensland entrance roads.

Eastern edge, General Purpose North 3, University of Queensland, St Lucia

As seen from informal courtyard; Duhig Tower beyond.

West elevation, General Purpose North 3, University of Queensland, St Lucia

Formal elevation facing University of Queensland entrance roads.

West elevation, General Purpose North 3, University of Queensland, St Lucia

Formal elevation facing University of Queensland entrance roads.

West elevation, General Purpose North 3, University of Queensland, St Lucia

Formal elevation facing University of Queensland entrance roads.

Prevalence of Parkinson’s disease in metropolitan and rural Queensland: A general practice survey

The prevalence of idiopathic Parkinson’s disease (IPD) in Australia is unclear. We estimated the prevalence of IPD, and other forms of parkinsonism, through the study of typical caseloads in general practice. A random sample of general practitioners (GPs) throughout Queensland (401 responses from 528 validated practice addresses) was asked to estimate the numbers of patients with IPD and parkinsonism seen in the preceding year. The estimated prevalence of diagnosed IPD in Queensland was 146 per 100 000 (95% CI = 136–155). A further 51 per 100 000 in the population were suspected by doctors to have IPD without formal diagnosis, whereas another 51 per 100 000 people may have non-idiopathic parkinsonism. Idiopathic Parkinson’s disease was more common in rural than metropolitan areas. Although most GPs were confident in making diagnoses of IPD, the majority had little or no confidence in their ability to treat the disease, especially in its later stages. Support from neurologists was perceived by GPs to be very good in cities, but poor in remote areas.

Breastfeeding and the responsibilities of GPs: a qualitative study of general practice registrars

The most recent National Health Survey reports that more than 80% of women initiate breastfeeding, while recent studies describe initiation rates of more than 90%. Yet fewer than 50% of women continue to breastfeed for 6 months or longer. This is at odds with National Health and Medical Research Council recommendations that 80% of infants be exclusively breastfed for the first 6 months of life. Women are more likely to initiate and continue to breastfeed if their doctor supports and encourages them to do so. Conversely, women perceive a neutral attitude by doctors toward breastfeeding to be similar to a negative attitude. Therefore, while doctors may not perceive their support or encouragement to be a determining factor in a woman’s breastfeeding decisions, women often place great emphasis on their GP's attitude to breastfeeding and are much more likely to think that information provided by a doctor is important. No previous research in Australia has addressed the issue of how GPs perceive their roles and responsibilities regarding breastfeeding. As part of a larger research project investigating the breastfeeding skills and knowledge of general practice registrars, this article reports the results of qualitative interviews with eight general practice registrars and their views and beliefs about GPs’ responsibilities to breastfeeding women.